This program is concerned with a study of the mechanism of citrate production by the prostate gland. The prostate has the unique function of accumulating and secreting extraordinarily high levels of citrate. This activity is required to maintain the normally high citrate content of semen although the function of citrate in the reproductive process (es) is unknown. The metabolic peculiarity of the prostate epithelium which accounts for this specialized activity is unresolved. Furthermore, alterations in the citrate metabolism of normal prostate appears to characterize metastic tissue arising from prostate carcinoma. This program will elucidate the biochemical activities of normal prostate metabolism of citrate. We will investigate the source of citrate (presumably from glucose via glycolysis and pyruvate formation) in rat ventral prostate. Also the mitochondrial synthesis of citrate as compared to mitochondrial citrate (via isocitrate) oxidation will be ascertained. By utilizing enzyme assay procedures and metabolic profiles we will determine if a deficient citrate oxidation is responsible for the high intracellular citrate content. We will also determine the compartmentalization of intracellular citrate in relation to phosphofructokinase activity. The activity of mitochondrial isocitrate dehydrogenase will be ascertained. The effects of androgen (testosterone) in regulating citrate production by prostate will be investigated. This supplement proposal requests the acquisition of a liquid scintillation counter and computer accessories required to perform the experiments, assays, and data processing.